In vitro analysis of glial cell function in ganglioside-deficient mice

Abstract

Gangliosides are a family of sialic acid-containing glycosphingolipids highly enriched in neuronal and glial membranes, where they play pleiotropic roles in nervous system function. In this glial cell biological study, we used mice deficient in glycosyltransferases involved in ganglioside biosynthesis to gain insights into the possible role of ganglioside overexpression or deficiency on glial cell proliferation, migration, and differentiation in vitro. Primary cultures of olfactory ensheathing cells, oligodendrocyte lineage cells, and Schwann cells isolated from beta1,4-N-acetylgalactosaminyl (beta1,4-GalNAc) transferase- and alpha-2,8-sialyltransferase-deficient mice demonstrated subtle differences in their behavior when compared with wild-type glia. Oligodendrocyte-axonal interactions were investigated in dissociated embryonic mixed spinal cord cultures in which axonal ensheathment with myelin internodes and organized nodes of Ranvier form. In these myelinating cultures, deficiency of complex gangliosides, as found in beta1,4-GalNAc T(-/-) mice, resulted in the temporal disorganization of K(v) and Na(+) channels at the nodes of Ranvier, similar to that seen in beta1,4-GalNAc T(-/-) mice in vivo. These data show that glycosyltransferase deficiency and the consequent ganglioside imbalance has subtle effects on a range of glial cell functions and that in vitro systems can be used to explore these in ways that complement whole animal physiology. Our results are also consistent with the absence of gross neurodevelopmental dysfunction in mice lacking a variety of different gangliosides, suggesting that ganglioside redundancy and substitution are mechanisms that compensate for the lack of a full complement of complex gangliosides.